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Molecular approach to annelid regeneration: cDNA subtraction cloning reveals various novel genes that are upregulated during the large‐scale regeneration of the oligochaete, Enchytraeus japonensis
Author(s) -
Myohara Maroko,
Niva Cintia Carla,
Lee Jae Min
Publication year - 2006
Publication title -
developmental dynamics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.634
H-Index - 141
eISSN - 1097-0177
pISSN - 1058-8388
DOI - 10.1002/dvdy.20849
Subject(s) - biology , annelid , blastema , suppression subtractive hybridization , gene , open reading frame , complementary dna , regeneration (biology) , genetics , cloning (programming) , cdna library , microbiology and biotechnology , anatomy , peptide sequence , computer science , programming language
To identify genes specifically activated during annelid regeneration, suppression subtractive hybridization was performed with cDNAs from regenerating and intact Enchytraeus japonensis , a terrestrial oligochaete that can regenerate a complete organism from small body fragments within 4–5 days. Filter array screening subsequently revealed that about 38% of the forward‐subtracted cDNA clones contained genes that were upregulated during regeneration. Two hundred seventy‐nine of these clones were sequenced and found to contain 165 different sequences (79 known and 86 unknown). Nine clones were fully sequenced and four of these sequences were matched to known genes for glutamine synthetase, glucosidase 1, retinal protein 4, and phosphoribosylaminoimidazole carboxylase, respectively. The remaining five clones encoded an unknown open‐reading frame. The expression levels of these genes were highest during blastema formation. Our present results, therefore, demonstrate the great potential of annelids as a new experimental subject for the exploration of unknown genes that play critical roles in animal regeneration. Developmental Dynamics 235:2051–2070, 2006. © 2006 Wiley‐Liss, Inc.